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Biomedical subjects

S Shiosaka

Publications and source records attributed to S Shiosaka.

At least 91 records · Page 5Linked to original sources

The presence of vasoactive intestinal polypeptide-like immunoreactive structures projecting from the myenteric ganglion of the stomach to the celiac ganglion revealed by a double-labelling technique.

The gastrofugal vasoactive intestinal polypeptide (VIP)-like immunoreactive (VIPI) structures in the rat were examined by the combined use of immunocytochemistry and retrograde tracing. Injection of biotin-wheat germ agglutinin into the celiac ganglion labeled many neurons in the myenteric ganglion of the stomach. Simultaneous staining with antiserum against VIP showed that some of these neurons are VIPI-positive. These findings indicate that VIPI neurons in the myenteric ganglion of the rat stomach project to the celiac ganglion.

Animals↗

A new method for producing a specific and high titre antibody against glutamate using colloidal gold as a carrier.

Antiserum against the glutamate was developed using colloidal gold as a carrier for the antigen. Glutamate was bound directly to colloidal gold without using cross-linking reagents such as glutaraldehyde, formaldehyde, or carbodiimide. Repeated injection of the colloidal gold-glutamate complex into the rabbit yielded a specific, high titre antiserum. Specificity was verified by radioimmunoassay and histochemically with absorption controls. The antiserum immunohistochemically detected numerous glutamate-containing neurons in the rat brain.

Animals↗

A neurotensin-immunoreactive pathway from the subiculum to the mammillary body in the rat.

The existence of a neurotensin-like immunoreactive (NT-IR) pathway from the subiculum to the mammillary body was demonstrated in very young rats using immunocytochemistry, knife cut, electric lesion and retrograde tracer techniques. Unilateral hemitransection of the dorsal fornix caused a marked reduction of NT-IR fibers in the ipsilateral mammillary body. Unilateral electric lesion of the subiculum also resulted in ipsilateral reduction of NT-IR fibers in the mammillary body. Biotin-wheat germ agglutinin (B-WGA), injected into the mammillary body, labeled neurons in the ipsilateral subiculum which were shown by double staining to have NT-IR cytoplasm. These findings demonstrate the presence of a major NT-IR fiber pathway from the subiculum to the mammillary body via the fornix.

Animals↗

Transplants in the peri- and intraventricular region grow better than those in the central parenchyma of the caudate.

The correlation between the growth of transplanted cells and behavior recovery in rats with lesions in the unilateral nigrostriatal dopamine (DA) pathway was investigated. Grafted cells in the lateral or third ventricle grew better than those in the caudate parenchyma. Immunocytochemical staining of tyrosine hydroxylase (TH) revealed that axons or dendrites of TH-like immunoreactive (TH-LI) cells transplanted periventricularly extended and ramified better than those of cells implanted in the central parenchyma of the caudate nucleus. The extent of ramification was well correlated with the recovery of behavioral imbalance. Data suggest that the growth and ramification of transplanted TH-LI cells are the most critical factors affecting behavior recovery, and this growth depends on the transplantation sites.

Animals↗

The formation of new neuronal circuit between transplanted nigral dopamine neurons and non-immunoreactive axon terminals in the host rat caudate nucleus.

Using immunoelectron microscopic techniques, whether or not host neuronal elements newly form synaptic contact with the grafted dopamine (DA) neurons in the caudate nucleus of the rat with unilateral lesion in the nigrostriatal DA pathway was examined. Tyrosine hydroxylase (TH) was used as a marker for DA-containing structures. Motor imbalances after the lesion and before or after the transplantation were assessed by the amount of circlings after the injection of Met-amphetamine. In animals which recovered well from motor imbalance, non-immunoreactive axon terminals made synaptic contact with grafted TH-positive cell bodies and their dendrites. Since the incidence of these synapses was quite low in poorly recovered animals, the formation of a new neuronal circuit may be one of the important bases for behavior recovery.

Animals↗

Distribution of calcitonin gene-related peptide-containing fibers in the urinary bladder of the rat and their origin.

By use of indirect immunofluorescence, this study demonstrated the presence of calcitonin gene-related peptide-like immunoreactive (CGRPI) fibers in the bladder of the rat. These fibers were abundant in the muscle layer, in which they ran parallel to the muscles, submucosa, and epithelium. No immunoreactive cells were detected. We also examined the origins of these fibers, using a method that combined biotinized retrograde tracer (biotin-wheat germ agglutinin) (B-WGA) and immunocytochemistry. Injection of the tracer into the bladder resulted in the demonstration of small to medium-sized labeled cells that contain CGRPI structures in single dorsal root ganglion cells mostly at the level of L6 and S1, but also a few at L2. Double-staining for CGRPI and immunoreactive P-like substance (SPI) indicated that there are cells in the dorsal root ganglia at the level of L6 and S1 that react to both, but that there are many CGRPI-positive cells that contain no demonstrable SPI; most of the latter are large.

Animals↗

The presence of a calcitonin gene-related peptide in the olivocochlear bundle in rat.

The origins of calcitonin gene-related peptide immunoreactive (CGRPI) fibers in the cochlea were examined in rats. Parasagittal transection of the brain just medial to the principal sensory trigeminal nucleus resulted in the ipsilateral disappearance of CGRPI fibers in the cochlea, indicating that the origins of these fibers lie in the central nervous system. Next, we used a highly sensitive method combining retrograde tracing and immunohistochemistry to identify the origins of the CGRPI fibers in the cochlea. After injection of biotin-wheat germ agglutinin (b-WGA) into the cochlea, CGRPI neurons in the ipsilateral lateral superior olivary nucleus also contained b-WGA granules. These findings indicated that CGRPI efferent fibers are major components of the olivocochlear bundle.

Animals↗

Sensitive double-labeling technique of retrograde biotinized tracer (biotin-WGA) and immunocytochemistry: light and electron microscopic analysis.

We developed a simple method to identify fiber projections that contain bioactive substances using biotinized wheat-germ agglutinin. Tracer that accumulated retrogradely in the soma was made visible under the light microscope by linking it with streptavidin-Texas red, which has a red fluorescence, while antigen was demonstrated in the same section by indirect immunofluorescence using fluorescein isothiocyanate with green fluorescence as a marker. Under the electron microscope, the tracer was identified using streptavidin-colloidal gold particles, while the antigen was demonstrated in the same ultrathin section using the peroxidase-antiperoxidase technique. This method makes it possible to demonstrate the locations of both the biotinized retrograde tracer and the antigen in the soma of the same neuron at both light and electron microscopic levels.

Animals↗

Cloning and characterization of a cDNA specific for bovine retina.

A retina-specific cDNA clone (pCR18) was selected from a bovine retinal cDNA library and characterized. The clone pCR18 consisted of 905 base pairs and hybridized to the mRNA of about 12S from the bovine retina, but not that from the brain or liver. The nucleotide sequence revealed a long open reading frame which encodes a 147 amino acid polypeptide of about 15,700 Da. No significant sequence homology with the predicted protein was found in the protein sequence library of about 3500. Messenger RNA which hybridized to pCR18 translated a polypeptide of about 19,000 Da in a reticulocyte translation system. Southern blot analysis indicated that the bovine genome contains a single copy of this gene. Furthermore, RNA dot analysis showed that the poly(A)+ RNA from the human retinoblastoma cell lines (Y79 and WERI) hybridized to pCR18, whose intensity was comparable to that of the bovine retina. In situ hybridization revealed that pCR18 was expressed mostly in some ganglion cells of the rat retina. The results suggest that cDNA clone (pCR18) encodes a protein specific for the retina and mRNA for pCR18 is mostly localized in the retinal ganglion cells and also expressed in the human retinoblastoma cells, although its function remains to be elucidated.

Animals↗

Distribution of neuropeptide Y in the forebrain and diencephalon: an immunohistochemical analysis.

The distribution of neuropeptide Y (NPY)-like immunoreactive (NPYI) structures in the rat forebrain and upper brainstem was examined by means of indirect immunofluorescence and peroxidase-antiperoxidase methods. The present study has demonstrated a much wider and more abundant distribution of NPYI structures in the forebrain and diencephalon than earlier studies in which antisera against avian pancreatic polypeptide or bovine pancreatic polypeptide were used, suggesting that NPY is involved in a variety of brain functions.

Animals↗

Calcitonin gene-related peptide projection from the ventromedial thalamic nucleus to the insular cortex: a combined retrograde transport and immunocytochemical study.

We employed a highly sensitive combination method of retrograde tracing and immunohistochemistry to identify calcitonin gene-related peptide (CGRP)-containing fiber pathways in the rat from the ventrolateral part of the ventrolateral thalamic nucleus (vl-vl) and the caudal continuation to the insular cortex. Biotin-wheat germ agglutinin (B-WGA) injected into the insular cortex labeled numerous neurons in the vl-vl and the caudal continuation ipsilaterally; simultaneous staining with CGRP antiserum revealed that some of these neurons are CGRP positive.

Animals↗

Immunohistochemical analysis of the cross-reaction of anti-rat histidine decarboxylase antibody with guinea-pig DOPA decarboxylase.

L-Histidine decarboxylase [L-histidine carboxylyase, HDC, EC 4.1.1.22] is an enzyme distinct from L-DOPA decarboxylase [L-aromatic amino acid carboxylyase, DDC, EC 4.1.1.28]: the two decarboxylases from fetal rat liver were completely separated from each other by DEAE-cellulose column chromatography and by affinity chromatography with L-carnosine as a ligand. The antibody raised against this HDC inhibited the HDC's from rat and guinea-pig brains very strongly, but their DDCs very weakly. However, in immunofluorescent histochemical studies, the antibody cross-reacted with DDC-like immunoreactive structures, such as chromaffin cells of the adrenal medulla, the raphe nucleus, the substantia nigra, and the locus coeruleus of the brain of guinea-pigs, but not of rats, suggesting that these two decarboxylases share some antigenic structures.

Adrenal Medulla↗

The descending alpha-MSHergic (alpha-melanocyte-stimulating hormone-ergic) projections from the zona incerta and lateral hypothalamic area to the inferior colliculus and spinal cord in the rat.

Neuronal pathways containing alpha-melanocyte-stimulating hormone (alpha-MSH) extending from the zona incerta and lateral hypothalamic area to the inferior colliculus and spinal cord were analyzed using both immunohistochemical localization and a retrograde tracer. Biotinized horseradish peroxidase injected into the inferior colliculus or the thoracic cord of the rat labeled a number of neurons in the zona incerta and lateral hypothalamic area. Simultaneous immunostaining of the same sections with alpha-MSH antiserum showed that some of these neurons are alpha-MSHergic.

Animals↗

Distribution of neuropeptide Y in the lower brainstem: an immunohistochemical analysis.

The distribution of neuropeptide Y (NPY)-like immunoreactive (NPYI) structures in the rat lower brainstem was examined by means of indirect immunofluorescence or peroxidase-antiperoxidase methods. In addition to the well known immunoreactive NPYI containing cell groups, the present study demonstrated a much wider distribution of immunoreactive NPYI cells in the lower brainstem, i.e. substantia nigra pars lateralis, interpeduncular nucleus, inferior colliculus, nucleus cuneiformis, dorsal tegmental nucleus of Gudden, nucleus laterodorsalis tegmenti, nucleus vestibularis medialis, nucleus vestibularis inferioris, in the fasciculus longitudinalis medialis and nucleus parvocellularis compacta. We also demonstrated an extensive network of NPYI fibers in various areas of the lower brainstem including the auditory system, viscerosensory system, visceromotor system, raphe nuclei, reticular formation, parabrachial area, locus coeruleus and interpeduncular nucleus, etc. The possible importance of NPY is briefly discussed.

Animals↗

Fine structure of peptidergic and catecholaminergic nerve fibers in the anterior cerebral artery and their interrelationship: an immunoelectron microscopic study.

This study shows (1) the ultrastructure of vasoactive intestinal polypeptide (VIP)-, substance P (SP)-, and neuropeptide Y (NPY)-containing nerve fibers in the walls of the cerebral arteries and (2) the relationship between these peptidergic (VIP, SP, and NPY) and catecholaminergic (CA) nerve terminals by immunohistochemistry combined with false transmitter (5-hydroxydopamine) histochemistry under the electron microscope. VIP-, SP-, and NPY-like immunoreactivity (VIPI, SPI, and NPYI) were found diffusely in the axoplasm and around the small clear vesicles in the nerve terminals. In a few cases, SPI was found within the large vesicles. Most of the VIPI terminals were ensheathed by the cytoplasm of the Schwann cells together with CA terminals, identified as those with a number of small granulated vesicles. In some cases, they were directly apposed to the smooth muscle cells at a distance of about 100 nm. SPI terminals were frequently solitary but about 30% were located together with CA and other (neither SPI nor CA) terminals ensheathed by Schwann cells, directly apposed to the smooth muscle cells at a distance of about 100 nm. On the other hand, NPYI terminals were also identified as CA terminals, indicating the coexistence of these two substances. These findings suggest a close interrelationship between peptidergic and CA nerve terminals in the neurogenic control of cerebral blood vessel function.

Animals↗

Coexistence of calcitonin gene-related peptide and substance P-like peptide in single cells of the trigeminal ganglion of the rat: immunohistochemical analysis.

The localization of calcitonin gene-related peptide (CGRP) and substance P (SP) in the rat trigeminal ganglion was examined by means of the indirect immunofluorescent method. About 40% of neurons in the ganglion contained CGRP-like immunoreactivity (CGRPI), while about 20% of neurons showed SP-like immunoreactivity (SPI). In serial sections, nearly all the SPI neurons contained CGRPI.

Animals↗